本文選題：線粒體　切入點(diǎn)：膜電位　出處：《華東師范大學(xué)》2011年碩士論文　論文類型：學(xué)位論文

【摘要】：線粒體是真核細(xì)胞中最重要的細(xì)胞器之一,除了為細(xì)胞提供能量,還參與細(xì)胞信號(hào)轉(zhuǎn)導(dǎo)、分化與生長(zhǎng)、凋亡等生命過程。許多疾病的發(fā)生與線粒體功能失常密切相關(guān),包括神經(jīng)退行性疾病、糖尿病、肥胖、腫瘤等。近年來,對(duì)線粒體功能的研究日益為研究人員所關(guān)注。因此,我們希望通過尋找靶向線粒體功能的溫和調(diào)節(jié)劑而獲得對(duì)糖脂代謝有改善作用的先導(dǎo)化合物。 線粒體膜電位是線粒體功能的一個(gè)重要指針,我們建立L6肌細(xì)胞線粒體膜電位調(diào)節(jié)劑高通量篩選模型,對(duì)國家新藥篩選中心小分子化合物庫近4000樣次化合物進(jìn)行隨機(jī)篩選,得到了一個(gè)低毒并且能夠濃度依賴的降低線粒體膜電位的小分子化合物L(fēng)GH00272。LGH00272可以比較溫和地降低L6肌細(xì)胞的線粒體膜電位,并且溫和的促進(jìn)L6肌細(xì)胞、大鼠原代肝細(xì)胞和分離的大鼠肝臟線粒體耗氧,同時(shí)對(duì)細(xì)胞中ATP的合成無明顯抑制,對(duì)乳酸的生成有輕微促進(jìn)。進(jìn)一步在代謝性細(xì)胞和整體動(dòng)物水平上評(píng)價(jià)該化合物的藥效表明：LGH00272能夠促進(jìn)L6肌細(xì)胞的葡萄糖吸收和脂肪酸氧化,降低HepG2肝癌細(xì)胞甘油三酯和膽固醇含量。在整體動(dòng)物實(shí)驗(yàn)中,LGH00272長(zhǎng)期給藥能夠使飲食誘導(dǎo)肥胖(diet induced obesity, DIO)小鼠的體重增長(zhǎng)更加緩慢,并且對(duì)攝食沒有影響。LGH00272還可以降低DIO小鼠血糖和膽固醇,腹腔注射葡萄糖耐量實(shí)驗(yàn)也進(jìn)一步證明LGH00272能夠提高DIO小鼠的糖耐受力。此外,LGH00272對(duì)糖異生也有明顯抑制,不僅能夠抑制大鼠原代肝細(xì)胞的糖輸出,還能降低長(zhǎng)期給藥DIO小鼠肝臟中糖異生關(guān)鍵基因的表達(dá)水平。LGH00272還降低了肝臟和脂肪比重,同時(shí)降低脂合成及增加脂代謝相關(guān)基因的表達(dá)。 綜上所述,我們通過線粒體膜電位高通量篩選得到了一個(gè)溫和的線粒體解偶聯(lián)劑,該化合物在細(xì)胞水平和整體動(dòng)物水平對(duì)糖脂代謝均有明顯的改善功能,結(jié)果表明通過尋找溫和的線粒體解偶聯(lián)劑可以獲得新穎的治療代謝綜合癥的先導(dǎo)化合物,對(duì)進(jìn)一步闡明線粒體和代謝綜合癥的關(guān)系起到積極的意義。
[Abstract]:Mitochondria is one of the most important organelles in eukaryotic cells. In addition to providing energy to the cells, mitochondria are also involved in cell signal transduction, differentiation and growth, apoptosis and other life processes. Many diseases are closely related to mitochondrial dysfunction. These include neurodegenerative diseases, diabetes, obesity, tumours, and so on. In recent years, the study of mitochondrial function has attracted more and more attention. We hope to obtain lead compounds that improve glucose and lipid metabolism by looking for mild regulators targeting mitochondrial function. Mitochondrial membrane potential is an important pointer to mitochondrial function. We established a high-throughput screening model for mitochondrial membrane potential modulators in L6 myocytes and randomly screened nearly 4000 small molecular compounds in the small molecular compound library of the National Center for New Drug screening. A low toxic and concentration-dependent low molecular compound, LGH00272.LGH00272, could decrease the mitochondrial membrane potential of L6 myocytes mildly, and promote L6 myocytes mildly. The primary rat hepatocytes and isolated rat liver mitochondria consumed oxygen, while the synthesis of ATP in the cells was not significantly inhibited. Further evaluation of the drug efficacy of the compound at the metabolic and overall animal levels showed that: LGH00272 promoted glucose absorption and fatty acid oxidation in L6 myocytes. Reduce the content of triglyceride and cholesterol in HepG2 hepatoma cells. In the whole animal experiment, the long-term administration of LGH00272 could slow the weight gain of diet-induced obese induced mice. LGH00272 also decreased blood glucose and cholesterol in DIO mice, and glucose tolerance test by intraperitoneal injection showed that LGH00272 could increase glucose tolerance in DIO mice. LGH00272 also inhibited glucose heterogenesis in DIO mice. It not only inhibited the glucose output of rat primary hepatocytes, but also decreased the expression level of glycosylated key genes in the liver of long-term DIO mice. LGH00272 also decreased the specific gravity of liver and fat. At the same time, it decreased lipid synthesis and increased the expression of genes related to lipid metabolism. In conclusion, we obtained a mild mtDNA uncoupling agent through high throughput screening of mitochondrial membrane potential, which can significantly improve glycolipid metabolism at cell level and animal level as a whole. The results show that a novel lead compound for the treatment of metabolic syndrome can be obtained by searching for mild mitochondrial uncoupling agents, which plays a positive role in further elucidating the relationship between mitochondria and metabolic syndrome.
【學(xué)位授予單位】：華東師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R341

【引證文獻(xiàn)】

相關(guān)博士學(xué)位論文 前1條

1 張哲;正電荷修飾的非標(biāo)記核酸熒光探針的研究及其在中藥抗腫瘤研究中的應(yīng)用[D];吉林大學(xué);2012年

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本文編號(hào)：1650628